Tryptase inhibitors

ABSTRACT

Compounds of the formula (I), in which M is a central building block selected from the group below in which Al, A2, K1 and K2 are as defined in the description, are novel effective tryptase inhibitors.

APPLICATION OF THE INVENTION

[0001] The invention relates to novel tryptase inhibitors which are usedin the pharmaceutical industry for preparing medicaments.

KNOWN TECHNICAL BACKGROUND

[0002] The international applications WO95/32945 (=U.S. Pat. No.5,656,660), WO96/09297 (=U.S. Pat. Nos. 6,022,969, 6,221,228),WO98/04537, WO99/12918, WO99/24395, WO99/24407, WO99/40073, WO99/40083(=DE19851299) and WO00/14097 describe low-molecular-weight bivalentcompounds for use as tryptase inhibitors.

DESCRIPTION OF THE INVENTION

[0003] It has now been found that the compounds of the formula I, whichare described in more detail below, have surprising and particularlyadvantageous properties. The invention provides compounds of the formulaI

[0004] in which

[0005] M is a central building block selected from the group below

[0006] A1 is -O-B1-A3-, -A5-B1-O-, —C(O)—, —C(O)—NH—, —NH—C(O)—,—O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or —O—C(O)—O—,

[0007] A2 is -O-B2-A4-, -A6-B2-O-, —C(O)—, —C(O)—NH—, —NH—C(O)—,—O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or —O—C(O)—O—,

[0008] A3 and A4 are identical or different and are —C(O)—, —C(O)—NH—,—NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or —O—C(O)—O—,

[0009] A5 and A6 are identical or different and are —C(O)—, —C(O)—NH—,—NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or —O—C(O)—O—,

[0010] B1 and B2 are identical or different and are 1-4C-alkylene,

[0011] K1 is -B3-X1, -B3-Y1 or -B3-Z1-B5-X1,

[0012] K2 is -B4-X2, -B4-Y2 or -B4-Z2-B6-X2,

[0013] B3 and B4 are identical or different and are a bond or1-4C-alkylene,

[0014] B5 and B6 are identical or different and are a bond or1-2C-alkylene,

[0015] X1 and X2 are Identical or different and are amino, aminocarbonylor amidino,

[0016] Y1 and Y2 are imidazol-1-yl,

[0017] Z1 and Z2 are identical or different and are 5,2-pyridinylene,6-methyl-5,2-pyridinylene, 4,1-piperidinylene, 3,6-indazolylene,3,6-indolylene, 1,3-phenylene, 1,4-phenylene, 1,3-cyclohexylene or1,4-cyclohexylene,

[0018] and where on the direct route between the terminal nitrogen atoms20 to 35, preferably 24 to 29, bonds have to be present,

[0019] and the salts of these compounds and the N-oxides and theirsalts, except for all those compounds in which one or more of thevariables B3, B4, B5 or B6 can assume the meaning of a bond, resultingin the direct linkage of two heteroatoms.

[0020] 1-4C-alkylene represents straight-chain or branched 1-4C-alkyleneradicals, for example the methylene [—CH₂—], ethylene [—CH₂—CH₂—],trimethylene [—CH₂—CH₂—CH₂—], tetramethylene [—CH₂—CH₂—CH₂—CH₂—],1,2-dimethyethylene [—CH(CH₃)—CH(CH₃) —], 1,1-dimethylethylene[—C(CH₃)₂—CH₂—], 2,2-dimethylethylene [—CH₂—C(CH₃)₂—], isopropylidene[—C(CH₃)₂—] or the 1-methylethylene [—CH(CH₃)—CH₂—] radical.

[0021] By definition, the groups Z1 and Z2 are located between groups B3and B5 (-B3-Z1-B5-) and B4 and B6 (-B4-Z2-B6-), respectively.Accordingly, in the divalent groupings mentioned by way of example (forexample 3,6-indolylene), the first number indicates the point ofattachment to the group B3 and B4, respectively, and the second numberindicates the point of attachment to the group B5 and B6, respectively.

[0022] The groups Z1 and Z2 may inter alia have the meanings1,4-cyclohexylene and 1,3-cyclohexylene. The invention embraces thecompounds of the formula I in which the groups B3, B5 and B4, B6,respectively, are attached (1e,4e), (1a,4a), (1e,4a), (1a,4e), (1e,3e),(1a,3a), (1e,3a) and (1a,3e) to the cyclohexylene radical. In thiscontext, particular preference is given to the (1e,4e) attachment (“e”means equatorial and “a” means axial).

[0023] In the context of this application, the term “terminal nitrogenatom” means in each case a nitrogen atom in the groupings designated X1,X2, Y1 and Y2.

[0024] If the group X1 or X2 contains only one nitrogen atom, thisnitrogen atom is the terminal nitrogen atom.

[0025] If the group X1 or X2 contains a plurality of nitrogen atoms, thenitrogen atom which is furthest from the atom by means of which the bondto the group B3 (B5) or B4 (B6) is established is the terminal nitrogenatom.

[0026] If the group Y1 or Y2 contains only one nitrogen atom, this ringnitrogen atom is the terminal nitrogen atom.

[0027] If the group Y1 or Y2 contains a plurality of nitrogen atoms, thering nitrogen atom which is furthest from the atom by means of which thebond to the group B3 or B4 is established is the terminal nitrogen atom.

[0028] According to the invention, the direct route between the nitrogenatoms which act as terminal nitrogen atoms in the groups defined as X1(Y1) or X2 (Y2) is considered to be the number of bonds which isobtained by counting the bonds which represent the shortest possibleconnection between the terminal nitrogen atoms.

[0029] The following example is meant to Illustrate the determination ofthe number of bonds on the direct route between two terminal nitrogenatoms:

[0030] Here, the direct route comprises 26 bonds.

[0031] The definitions of M contain chemical formulae, such as, forexample,

[0032] This formula represents 2,6-bis(prop-1-ynyl)pyridine,2,5-bis(prop-1-ynyl)pyridine, 2,4-bis(prop-1ynyl)pyridine,2,3-bis(prop-1-ynyl)pyridine, 3,6-bis(prop-1-ynyl)pyridine,3,5-bis(prop-1-ynyl)pyridine or 3,4-bis(prop-1-ynyl)pyridine. Thisapplies correspondingly to the other chemical formulae given in thedefinitions of M.

[0033] Preference is given to compounds of the formula I whose molecularweight is below 600 g/mol.

[0034] Suitable salts for compounds of the formula I are all acidaddition salts. Particular mention may be made of the pharmaceuticallyacceptable salts of inorganic and organic acids customarily used inpharmacy. Those suitable are water-soluble and water-insoluble acidaddition salts with acids such as, for example, hydrochloric acid,hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid, aceticacid, citric acid, D-gluconic acid, benzoic acid,2-(4-hydroxybenzoyl)benzoic acid, butyric acid, sulfosalicylic acid,maleic acid, lauric acid, malic acid, fumaric acid, succinic acid,oxalic acid, tartaric acid, embonic acid, stearic acid, toluenesulfonicacid, mehethanesulfonic acid or 3-hydroxy-2-naphthoic acid, where theacids are employed in salt preparation—depending on whether a mono- orpolybasic acid is concerned and depending on which salt is desired-in anequimolar quantitative ratio or one differing therefrom.

[0035] Pharmacologically unacceptable salts which can be obtainedinitially as process products, for example in the preparation of thecompounds according to the invention on an industrial scale, areconverted into pharmacologically acceptable salts by processes known tothe person skilled in the art.

[0036] It is known to the person skilled in the art that the compoundsaccording to the invention, and also their salts, may contain varyingamounts of solvents, for example when they are isolated in crystallineform. The invention therefore also embraces all solvates and inparticular all hydrates of the compounds of the formula I, and also allsolvates and in particular all hydrates of the salts of the compounds ofthe formula I.

[0037] Compounds of the formula I which are to be emphasized are thosein which M is a central building block selected from the group below

[0038] A1 is -O-B1-A3-, -A5-B1-O-, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—,—NH—C(O)—NH— or —O—C(O)—O—,

[0039] A2 is -O-B2-A4-, -A6-B2-O-, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—,—NH—C(O)—NH— or —O—C(O)—O—,

[0040] A3 and A4 are identical or different and are —C(O)—NH—,—NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or —O—C(O)—O—,

[0041] A5 and A6 are identical or different and are —NH—C(O)—,—O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or —O—C(O)—O—,

[0042] B1 and B2 are identical or different and 1-2C-alkylene,

[0043] K1 is -B3-Z1-B5-X1,

[0044] K2 is -B4-Z2-B6-X2,

[0045] B3 and B4 are identical or different and are a bond or1-2C-alkylene,

[0046] B5 and B6 are identical or different and are a bond or1-2C-alkylene,

[0047] X1 and X2 are identical or different and are amino or amidino,

[0048] Z1 and Z2 are identical or different and are 1,3-phenylene,1,4-phenylene, 1,3-cyclohexylene or 1 ,4-cyclohexylene,

[0049] and where on the direct route between the terminal nitrogen atoms20 to 35, preferably 24 to 29, bonds have to be present,

[0050] and the salts of these compounds, the N-oxides and their salts.

[0051] Compounds of the formula I which are to be particularlyemphasized are those in which M is a central building block selectedfrom the group below

[0052] A1 is —O—C(O)—NH—,

[0053] A2 is —O—C(O)—NH—,

[0054] K1 is -B3-Z1-B5-X1,

[0055] K2 is -B4-Z2-B6-X2,

[0056] B3 and B4 are identical and are methylene,

[0057] B5 and B6 are identical and are methylene,

[0058] X1 and X2 are identical and are amino,

[0059] Z1 and Z2 are identical or different and are 1,3-phenylene or1,4-phenylene,

[0060] and the salts of these compounds, the N-oxides and their salts.

[0061] Preferred compounds of the formula I are2,6-bis-[3-(3-aminomethylbenzylaminocarbonyloxy)prop-1-ynyl]pyridine and

[0062]2,6-bis-[3-(4-aminomethylbenzylaminocarbonyloxy)prop-1-ynyl]pyridine

[0063] and the salts of these compounds, the N-oxides and their salts.

[0064] The compounds of the formula I are constructed of a large numberof building blocks (M, A1, A2, A3, A4, A5, A6, B1, B2, B3, B4, B5, B6,X1, X2, Y1, Y2, Z1 and Z2). In principle, they can be synthesizedstarting with any of these building blocks. If the compounds of theformula I are constructed largely symmetrically, it is favorable tostart the synthesis with the central building block M, whereas in thecase of predominantly asymmetrical compounds of the formula I asynthesis starting with one of the end groups K1 or K2 may beadvantageous.

[0065] Suitable starting materials for synthesizing the compounds of theformula I according to the invention are, for example,2,6-dibromopyridine, 2,5-dibromopyridine, 3,5-dibromopyridine,2,5-dibromopyridazine, 2,4-dibromopyrimidine,2,4-dibromo-[1,3,5]-triazine or 1,3-dihydroxybenzene.

[0066] Here, the building blocks are linked using always the samepattern, known per se to the person skilled in the art.

[0067] It is known to the person skilled in the art that the compoundsof the formula I can either be synthesized building block by buildingblock, or by Initially constructing relatively large fragmentsconsisting of several individual building blocks, which can then bejoined to give the complete molecule.

[0068] Owing to the meanings which the individual building blocks of thecompounds of the formula I can assume, ether [—O—], keto [—C(O)—], amide[—C(O)—NH—, —NH—C(O)—], carbamate [—NH—C(O)—O—, —O—C(O)NH—], carbamide[—NH—C(O)—NH—] or carbonate [—O—C(O)—O—] bridges can be present in thecompounds of the formula I.

[0069] How to prepare such bridges is known per se to the person skilledin the art; suitable methods and starting materials for theirpreparation are described, for example, in March, Advanced OrganicChemistry, Reactions, Mechanisms and Structure, Third Edition, 1985,John Wiley & Sons.

[0070] Ether bridges can be prepared, for example, by the method ofWilliamson.

[0071] Keto bridges can be introduced, for example, as a component ofrelatively large building blocks, such as, for example, carboxylic acidderivatives.

[0072] There is also a large number of known methods for preparing amidebridges. An example which may be mentioned here is the reaction of acylchlorides with primary or secondary amines. Furthermore, reference isalso made to all the methods which have been developed for peptidechemistry.

[0073] Carbamate bridges can be prepared, for example, by reactingchloroformates with amines. The chloroformates for their part can besynthesized from alcohols and phosgene. A further variant forconstructing carbamate bridges is the addition of alcohols toisocyanates. Similarly to carbamate bridges, it is possible to preparecarbonate bridges starting from chloroformates, by reaction withalcohols (instead of amines).

[0074] Carbamide bridges can be prepared, for example, by reactingisocyanates with amines.

[0075] The preparation of the compounds of the formula I may be shown inan exemplary manner using the reaction schemes below. Reaction scheme 1shows the preparation of the exemplary compounds 1 and 2. Othercompounds of the formula I can be prepared analogously, or by using theabovementioned methods known per se to the person skilled in the art.

[0076] It is also possible to convert compounds of the formula I byderivatization into other compounds of the formula I. Thus, for example,compounds of the formula I having a nitrogen-containing heteroaryl,heteroarylene or heterocycloalkylene building block can be converted byoxidation into the corresponding N-oxides.

[0077] The N-oxidation is carried out in a manner which is likewiseknown to the person skilled in the art, for example using hydrogenperoxide in methanol or m-chloroperoxybenzoic acid in dichloromethane atroom temperature. Which reaction conditions are required in theparticular case for carrying out the process is known to the personskilled In the art owing to his expert knowledge.

[0078] It is furthermore known to the person skilled in the art that ifthere are a number of reactive centers of a starting material orintermediate, it may be necessary to block one or more reactive centerstemporarily by protective groups in order to allow a reaction to proceedspecifically at the desired reaction center. A detailed description ofthe use of a large number of proven protective groups is found, forexample, in T.W. Greene, Protective Groups in Organic Synthesis, JohnWiley & Sons, 1991.

[0079] The isolation and purification of the substances according to theinvention is carried out in a manner known per se, for example bydistilling off the solvent under reduced pressure and recrystallizingthe resulting residue from a suitable solvent or subjecting It to one ofthe customary purification methods, such as, for example, columnchromatography on a suitable support material.

[0080] Salts are obtained by dissolving the free compound in a suitablesolvent (for example a ketone, such as acetone, methyl ethyl ketone ormethyl isobutyl ketone, an ether, such as diethyl ether, tetrahydrofuranor dioxane, a chlorinated hydrocarbon, such as methylene chloride orchloroform, or a low-molecular-weight aliphatic alcohol, such as ethanolor isopropanol) which contains the desired acid or base, or to which thedesired acid or base is then added. The salts are obtained by filtering,reprecipitating, precipitating with a nonsolvent for the addition saltor by evaporating the solvent. Salts obtained can be converted byalkalization or by acidification into the free compounds, which in turncan be converted into salts. In this way, pharmacologically unacceptablesalts can be converted into pharmacologically acceptable salts.

[0081] The examples below serve to illustrate the invention in moredetail without restricting it. Likewise, further compounds of theformula I, whose preparation is not explicitly described, can beprepared in an analogous manner or in a manner familiar per se to theperson skilled in the art using customary process techniques.

[0082] In the examples below, the abbreviation RT stands for roomtemperature, calc. for calculated and MS for mass spectrometry. Thecompounds mentioned by way of example and their salts are the preferredsubject of the invention.

EXAMPLES

[0083] End Products

[0084] General Procedure

[0085] A solution of the Boc-protected divalent compound (A1, A2; 1.0mmol) in question in CH₂Cl₂ (2 ml) is admixed with trifluoroacetic acid(14 ml) and stirred at RT for 1 h. A saturated solution of HCl indiethyl ether (10 ml) is then added dropwise to the reaction mixture.The resulting precipitate is immediately filtered off and washed withdiethyl ether (3×5 ml). Drying under reduced pressure gives the titlecompounds (end products 1 and 2) as colorless solids.

[0086] 1.2,6-Bis-[3-(3-aminomethylbenzylaminocarbonyloxy)prop-1-ynyl]pyridinetrihydrochloride

[0087] MS: calc.: C₂₉H₂₉N₅O₄ (511.6), found: [MH⁺] 512.3

[0088]2.2,6-Bis-[3-(4-aminomethylbenzylaminocarbonyloxy)prop-1-ynyl]pyridinetrihydrochloride

[0089] MS: calc.: C₂₉H₂₉N₅O₄ (511.6), found: [MH⁺] 512.3

[0090] Starting Materials

[0091] A1.2,6-Bis-[3-(3-N-tert-butoxycarbonylaminomethylbenzylaminocarbonyloxy)prop-1ynyl]pyridine

[0092] N,N-carbonyldiimidazole (1.04 g, 8.15 mmol) is added to asolution of 2,6-bis-(3-hydroxyprop-1ynyl)pyridine (A3, 0.5 g, 2.7 mmol)in absolute CH₂Cl₂ (12 ml), and the mixture is stirred at RT for 2.5 h.The reaction solution is diluted with CH₂Cl₂ (12 ml) and extracted witha semisaturated aqueous NaCl solution (25 ml). The organic phase isdried over MgSO₄, filtered off and concentrated under reduced pressure.The resulting residue is taken up in absolute CH₂Cl₂ (12 ml),3-(tert-butyloxycarbonylaminomethyl)benzylamine (1.28 g, 5.4 mmol) isadded and the mixture is stirred at RT overnight. The reaction solutionis diluted with CH₂Cl₂ (12 ml) and extracted with a semisaturatedaqueous NaCl solution (25 ml). The organic phase is dried over MgSO₄,filtered off and concentrated under reduced pressure. Furtherpurification is carried out by chromatography [toluene/acetone(7.5:2.5)] on a silica gel column. This gives the title compound (1.0 g)as a colorless solid. TLC, silica gel (glass plates), [toluene/acetone(7:3)], R_(f)=0.47.

[0093] MS: calc.: C₃₉H₄₅N₅O₈ (711.8), found: [MH⁺] 712.0; [MNa⁺] 734.3

[0094] A2.2,6-Bis-[3-(4-N-tert-butoxycarbonylaminomethylbenzylaminocarbonyl-3-oxy)prop-1-ynyl]pyridine

[0095] N,N-carbonyldiimidazole (1.04 g, 8.15 mmol) is added to asolution of 2,6-bis-(3-hydroxyprop-1ynyl)pyridine (A3, 0.5 9, 2.7 mmol)in absolute CH₂Cl₂ (12 ml), and the mixture is stirred at RT for 2.5 h.The reaction solution is diluted with CH₂Cl₂ (12 ml) and extracted witha semisaturated aqueous NaCl solution (25 ml). The organic phase isdried over MgSO₄, filtered off and concentrated under reduced pressure.The resulting residue is taken up in absolute CH₂Cl₂ (12 ml),4-(tert-butyloxycarbonylaminomethyl)benzylamine (1.28 g, 5.4 mmol) isadded and the mixture is stirred at RT overnight. Diethyl ether (20 ml)is added to the reaction solution and the resulting precipitate isfiltered off, washed with diethyl ether (15 ml) and dried. This givesthe title compound (1.0 g) as a colorless solid. TLC, silica gel (glassplates), [toluene/acetone (7:3)], R_(f)=0.43.

[0096] MS: calc.: C₃₉H₄₅N₅O₈ (711.8), found: [MH⁺] 712.0; [MNa+] 734.3

[0097] A3. 2,6-Bis-(3-hydroxyprop-1-ynyl)pyridine

[0098] Pd(Ph₃P)₄ (99 mg, 2%), CuBrSMe₂ (38 mg) and propargyl alcohol(1.3 ml, 21 mmol) are added successively to a solution of2,6-dibromopyridine (5.0 g, 21.1 mmol) in triethylamine (120 ml), andthe mixture is stirred at RT for 10 min and then under reflux at 80° C.for 2 h. After cooling, the reaction mixture is filtered off throughkieselguhr, and the filter cake is washed with ethyl acetate (20 ml).The organic phase is concentrated under reduced pressure. Furtherpurification is carried out by chromatography [toluene/acetone (7:3)] ona silica gel column. This gives the title compound (2.65 g) as acolorless solid. TLC, silica gel (glass plates) [toluene/acetone (7:3)],R_(f)=0.15.

[0099] MS: calc.: C₁₁H₉NO₂ (187.2), found: [MH⁺] 188.0

[0100] Commercial Utility

[0101] As tryptase inhibitors, the compounds according to the inventionhave useful pharmacological properties which make them commerciallyutilizable. Human tryptase is a serin protease which is the main proteinin human mast cells. Tryptase comprises eight closely related enzymes(α1, α2, β1a, β1b, β2, β3, mMCP-7-like-1, mMCP-7-like-2; 85 to 99%sequence identity) (cf. Miller et al., J. Clin. Invest. 84 (1989)1188-1195; Miller et al., J. Clin. Invest. 86 (1990) 864-870;Vanderslice et al., Proc. Natl. Acad. Sci., USA 87 (1990) 3811-3815;Pallaoro et al., J. Biol. Chem. 274 (1999) 3355-3362). However, only theβ-tryptases (Schwartz et al., J. Clin. Invest. 96 (1995) 2702-2710;Sakai et al., J. Clin. Invest. 97 (1996) 988-995) are activatedintracellularly and stored in catalytically active form in secretorygranules. Compared with other known serin proteases, such as, forexample, trypsin or chymotrypsin, tryptase has some special properties(Schwartz et al., Methods Enzymol. 244, (1994), 88-100; G. H. Caughey,“Mast cell proteases in immunology and biology”. Marcel Dekker, Inc.,New York, 1995). Tryptase from human tissue has a noncovalently-linkedtetrameric structure which has to be stabilized by heparin or otherproteoglycanes to be proteolytically active. Together with otherinflammatory mediators, such as, for example, histamine andproteoglycanes, tryptase is released when human mast cells areactivated. Because of this, tryptase is thought to play a role in anumber of disorders, in particular in allergic and inflammatorydisorders, firstly because of the importance of the mast cells in suchdisorders and secondly since an increased tryptase concentration wasobserved in a number of disorders of this type. Thus, tryptase isassociated, inter alia, with the following diseases: acute and chronic(in particular inflammatory and allergen-induced) airway disorders ofvarious origins (for example bronchitis, allergic bronchitis, bronchialasthma, COPD); interstitial lung disorders; disorders based on allergicreactions of the upper airways, (pharynx, nose) and the adjacent regions(for example paranasal sinuses, conjunctivae), such as, for example,allergic conjunctivitis and allergic rhinitis; disorders of thearthritis type (for example rheumatoid arthritis); autoimmune disorders,such as multiple sclerosis; furthermore neurogenic inflammations,arteriolosclerosis and cancer; moreover periodontitis, anaphylaxis,interstitial cystitis, dermatitis, psoriasis, sclerodermia/systemicsclerosis, inflammatory intestinal disorders (Crohn's disease,inflammatory bowel disease) and others. In particular, tryptase seems tobe connected directly to the pathogenesis of asthma (Caughey, Am. J.Respir. Cell Mol. Biol. 16 (1997), 621-628; R. Tanaka, “The role oftryptase in allergic inflammation” in: Protease Inhibitors, IBC LibrarySeries, 1979, Chapter 3.3.1-3.3.23).

[0102] A further subject of the invention are the compounds according tothe invention for use in the treatment and/or prophylaxis of diseases,in particular the diseases mentioned.

[0103] The invention likewise relates to the use of the compoundsaccording to the invention for preparing medicaments which are employedfor the treatment and/or prophylaxis of the diseases mentioned.

[0104] Medicaments for the treatment and/or prophylaxis of the diseasesmentioned, which contain one or more of the compounds according to theinvention, are furthermore a subject of the invention.

[0105] The medicaments are prepared by processes which are known per seand are familiar to the person skilled in the art. As medicaments, thecompounds according to the invention (=active compounds) are eitheremployed as such, or preferably in combination with suitablepharmaceutical excipients, for example in the form of tablets, coatedtablets, capsules, suppositories, patches, emulsions, suspensions, gelsor solutions, the active compound content advantageously being between0.1 and 95%.

[0106] The person skilled in the art is familiar on the basis of his/herexpert knowledge with the excipients which are suitable for the desiredpharmaceutical formulations. In addition to solvents, gel-formingagents, ointment bases and other active compound vehicles, it ispossible to use, for example, antioxidants, dispersants, emulsifiers,preservatives, solubilizers or permeation promoters.

[0107] For the treatment of diseases of the respiratory tract, thecompounds according to the invention are preferably also administered byinhalation, preferably in the form of an aerosol, with the aerosolparticles of solid, liquid or mixed composition having a diameter offrom 0.5 to 10 μm, advantageously of from 2 to 6 μm.

[0108] The aerosol can be produced, for example, using pressure-drivennozzle nebulizers or ultrasonic nebulizers, advantageously, however,using propellant gas-driven metered aerosols or by means of thepropellant gas-free use of micronized active compounds from inhalationcapsules.

[0109] Depending on the inhalation system employed, the administrationforms also contain, In addition to the active compounds, the requisiteauxiliary substances, for example propellant gases (e.g. Frigen in thecase of metered aerosols), surface-active substances, emulsifiers,stabilizers, preservatives, aromatizing agents, fillers (e.g. lactose inthe case of powder inhalers) and, where appropriate, additional activecompounds.

[0110] For the purposes of inhalation, a large number of appliances areavailable which can be used to generate aerosols of optimal particlesize and administer them using an inhalation technique which is asappropriate as possible for the patient. In addition to usingattachments (spacers and expanders) and pear-shaped containers (e.g.Nebulator® and Volumatic®), and also automatic spray puff releasers(Autohaler®) for metered aerosols, a number of technical solutions areavailable, particularly in the case of the powder inhalers (e.g.Diskhaler®, Rotadisk®, Turbohaler® or the inhaler described in Europeanpatent application 0 505 321), which technical solutions can be used toachieve optimal administration of the active compound.

[0111] For the treatment of dermatoses, the compounds according to theinvention are in particular used In the form of those medicaments whichare suitable for topical administration. For the preparation of themedicaments, the compounds according to the invention (=activecompounds) are preferably mixed with suitable pharmaceutical excipientsand further processed to give suitable pharmaceutical formulations.Suitable pharmaceutical formulations which may be mentioned are, forexample, powders, emulsions, suspensions, sprays, oils, ointments, fattyointments, creams, pastes, gels or solutions.

[0112] The medicaments according to the invention are prepared byprocesses known per se. The dosage of the active compounds in the caseof systemic therapy (p.o. or i.v.) is between 0.1 and 10 mg per kilogramper day.

[0113] Biological investigations

[0114] The documented pathophysiological effects of mast cell tryptaseare caused directly by the enzymatic activity of the protease.Accordingly, they are reduced or blocked by inhibitors which inhibit theenzymatic activity of the tryptase. A suitable measure for the affinityof a reversible inhibitor to the target protease is the equilibriumdissociation constant K_(i) of the enzyme-inhibitor complex. This K_(i)value can be determined via the effect of the inhibitor on thetryptase-induced cleavage of a chromogenic peptide-p-nitroanilidesubstrate or a fluorogenic peptide-aminomethylcoumarin substrate.

[0115] Methodology

[0116] The dissociation constants for the tryptase-inhibitor complexesare determined under equilibrium conditions in accordance with thegeneral proposals of Bieth (Bieth JG, Pathophysiological Interpretationof kinetic constants of protease inhibitors, Bull. Europ. Physiopath.Resp. 16:183-195, 1980) and the methods of Sommerhoff et al. (SommerhoffCP et al., A Kazal-type inhibitor of human mast cell tryptase: Isolationfrom the medical leech Hirudo medicinalis, characterization, andsequence analysis, Biol. Chem. Hoppe-Seyler 375: 685-694, 1994).

[0117] Human tryptase is isolated from lung tissue or preparedrecombinantly; the specific activity of the protease, determined bytitration, is usually greater than 85% of the theoretical value. In thepresence of heparin (0.1-50 μg/ml) for stabilizing the protease,constant amounts of the tryptase are incubated with increasing amountsof the inhibitors. After an equilibrium between the reaction partnershas formed, the remaining enzyme activity after addition of thepeptide-p-nitroanilide substrate tos-GlyPro-arg-pNA is determined andthe cleavage of the latter is monitored at 405 nm for 3 min.Alternatively, the remaining enzymatic activity can also be determinedusing fluorogenic substrates. The apparent dissociation constantsK_(iapp) (i.e. in the presence of substrate) are subsequently determinedby adapting the enzyme rates to the general equation for reversibleinhibitors (Morrison JF, Kinetics of the reversible inhibition ofenzyme-catalyzed reactions by tight-binding inhibitors, Biochim.Biophys. Acta 185,269-286,1969) using non-linear regression:

V ₁/V₀ =1−{E _(t) +I _(t) +K _(iapp)[(E _(t) +l _(t) +K _(iapp))²−4E_(t) l _(t)]^(1/2)}/2E _(t)

[0118] V₁ and V₀ are the rates in the presence and absence,respectively, of the inhibitor, and E_(t) and I_(t) are the tryptase andinhibitor concentrations, respectively.

[0119] The apparent dissociation constants determined for the compoundsaccording to the invention are shown in Table A below, where the numbersof the compounds correspond to the numbers of the compounds in theexamples. TABLE A Inhibition of human tryptase Compound K_(lapp) (μM) 10.0026 2 0.006

1. A compound of the formula I

in which M is a central building block selected from the group below

A1 is -O-B1-A3-, -A5-B1-O-, —C(O)—, —C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—,—NH—C(O)—O—, —NH—C(O)—NH— or —O—C(O)—O—, A2 is -O-B2-A4-, -A6-B2-O-,—C(O)—, —C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or—O—C(O)—O—, A3 and A4 are identical or different and are —C(O)—,—C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or—O—C(O)—O—, A5 and A6 are identical or different and are —C(O)—,—C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or—O—C(O)—O—, B1 and B2 are identical or different and are 1-4C-alkylene,K1 is -B3-X1, -B3-Y1 or -B3-Z1-B5-X1, K2 is -B4-X2, -B4-Y2 or-B4-Z2-B6-X2, B3 and B4 are identical or different and are a bond or1-4C-alkylene, B5 and B6 are identical or different and are a bond or1-2C-alkylene, X1 and X2 are identical or different and are amino,aminocarbonyl or amidino, Y1 and Y2 are imidazol-1-yl, Z1 and Z2 areidentical or different and are 5,2-pyridinylene,6-methyl-5,2-pyridinylene, 4,1-piperidinylene, 3,6-indazolylene,3,6-indolylene, 1,3-phenylene, 1,4-phenylene, 1,3-cyclohexylene or1,4cyclohexylene, and where on the direct route between the terminalnitrogen atoms 20 to 35 bonds have to be present, and the salts of thiscompound and the N-oxides and their salts, except for all thosecompounds in which one or more of the variables B3, B4, B5 or B6 canassume the meaning of a bond, resulting in the direct linkage of twoheteroatoms.
 2. A compound of the formula 1 as claimed in claim 1 inwhich M is a central building block selected from the group below

A1 is -O-B1-A3-, -A5-B1-O-, —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—,—NH—C(O)—NH— or —O—C(O)—O—, A2 is -O-B2-A4-, -A6-B2-O-, —NH—C(O)—,—O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or —O—C(O)—O—, A3 and A4 areidentical or different and are —C(O)—NH—, —NH—C(O)—, —O—C(O)—NH—,—NH—C(O)—O—, —NH—C(O)—NH— or —O—C(O)—O—, A5 and A6 are identical ordifferent and are —NH—C(O)—, —O—C(O)—NH—, —NH—C(O)—O—, —NH—C(O)—NH— or—O—C(O)—O—, B1 and B2 are identical or different and 1-2C-alkylene, K1is -B3-Z1-B5-X1, K2 is -B4-Z2-B6-X2, B3 and B4 are identical ordifferent and are a bond or 1-2C-alkylene, B5 and B6 are identical ordifferent and are a bond or 1-2C-alkylene, X1 and X2 are identical ordifferent and are amino or amidino, Z1 and Z2 are identical or differentand are 1,3-phenylene, 1,4-phenylene, 1,3-cyclohexylene or 1,4-cyclohexylene, and where on the direct route between the terminalnitrogen atoms 20 to 35 bonds have to be present, and the salts of thiscompound, the N-oxides and their salts.
 3. A compound of the formula Ias claimed in claim 1 in which M is a central building block selectedfrom the group below

A1 is —O—C(O)—NH—, A2 is —O—C(O)—NH—, K1 is -B3-Z1-B5-X1, K2 is-B4-Z2-B6-X2, B3 and B4 are identical and are methylene, B5 and B6 areidentical and are methylene, X1 and X2 are identical and are amino, Z1and Z2 are identical or different and are 1,3-phenylene or1,4-phenylene, and the salts of this compound, the N-oxides and theirsalts.
 4. A compound of the formula I as claimed in claim 1 having thechemical name2,6-bis-[3-(3-aminomethylbenzylaminocarbonyloxy)prop-1-ynyl]pyridine or2,6-bis-[3-(4-aminomethylbenzylaminocarbonyloxy)prop-1-ynyl]pyridine andthe salts of this compound, the N-oxides and their salts.
 5. A compoundof the formula I as claimed in claim 1 or 2 where on the direct routebetween the terminal nitrogen atom 24 to 29 bonds are present.
 6. Acompound of the formula I as claimed in any of claim 1, 2 or 3 having amolecular weight of below 600 g/mol.
 7. A compound of the formula I asclaimed in claim 1 for treating diseases.
 8. A medicament comprising oneor more compounds of the formula I as claimed in claim 1 together withthe customary pharmaceutical auxiliaries and/or excipients.
 9. The useof compounds of the formula I as claimed in claim 1 for preparingmedicaments for treating respiratory disorders.